Objective · To analyze target genes of human miR-223-3p (hsa-miR-223-3p) and their biological processes, and explore biomarkers related to diabetes mellitus (DM). Methods · starBase database was used to screen hsa-miR-223-3p target genes, and Gene Ontology (GO) enrichment analysis, Kyoto Encyclopedia of Genes and Genome (KEGG) and Reactome pathway analysis were performed. Hub genes were obtained by constructing protein-protein interaction (PPI) network, and the most significant modules were screened out. The hub genes related to DM were analyzed by Venn diagram. Results · A total of 870 hsa-miR-223-3p target genes were screened out. The GO enrichment analysis, KEGG and Reactome pathway analysis showed that the target genes were mainly related to the regulation of RNA polymerase Ⅱ promoter, cell response to insulin stimulation, RNA binding, etc, and were mainly enriched in insulin secretion, ubiquitin-mediated proteolysis and estrogen-dependent gene expression. There were 31 hub genes in PPI network, and hub gene PRKACB participated in insulin secretion pathway. Top 3 modules were identified as follows: module 1 was involved in ubiquitin-mediated proteolysis, module 2 was involved in RNA transport and cell cycle, and module 3 was involved in endocytosis. Conclusion · Hsa-miR-223-3p may participate in a variety of biological processes through target genes, and the hub gene PRKACB may provide assistance for exploring the pathogenesis of DM.